Designing of the expressing eukaryotic plasmid of the G1 epitope of bovine ephemeral fever virus G glycoprotein in human embryonic kidney cells

Pasandideh, Reza; Beigi Nasiri, Mohamad Taghi; Seifiabad Shapouri, Masoud Reza; Fayazi, Jamal; Roshanfekr, Hedayatallah; Lotfi, Mohsen

doi:10.22055/ivj.2017.60443.1793

Designing of the expressing eukaryotic plasmid of the G1 epitope of bovine ephemeral fever virus G glycoprotein in human embryonic kidney cells

Document Type : Research Paper

Authors

Reza pasandideh ¹

Mohamad Taghi Beigi Nasiri ²

Masoud Reza Seifiabad Shapouri ³

Jamal Fayazi ⁴

Hedayatallah Roshanfekr ²

Mohsen Lotfi ⁵

¹ PhD Student of Animal Genetics and Breeding, Faculty of Animal and Food Technology Ramin Agriculture and Natural Resources University of Khuzestan, Iran

² Professor, Department of Animal Science, Faculty of Animal and Food Technology Ramin Agriculture and Natural Resources University of Khuzestan, Iran

³ Professor, Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

⁴ Associate Professor, Department of Animal Science, Faculty of Animal and Food Technology Ramin Agriculture and Natural Resources University of Khuzestan, Iran

⁵ Research Assistant Professor, Department of Quality Control of Biological Products, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension, Karaj, Iran

10.22055/ivj.2017.60443.1793

Abstract

The G glycoprotein of bovine ephemeral fever virus (BEFV) has been identified as a plausible candidate for immunization against BEF disease. In recent years, this protein has been investigated to produce a recombinant vaccine. The aim of the present study was to construct a eukaryotic plasmid, expressing G1 epitope of BEFV G glycoprotein gene, for application as a possible DNA vaccine in future studies. For this purpose, the G1 epitope of G glycoprotein gene was cloned in a eukaryotic expression vector, pEGFP-N1, under the control of the human cytomegalovirus (CMV) promoter. Then, the recombinant pEGFPN1-G1 construct was transfected into human embryonic kidney (HEK 293) cell line and the expression efficiency was verified by indirect immunoﬂuorescent (IFA) staining and immunoblotting techniques. Observation of intracytoplasmic ﬂuorescence in transfected cells and appearance of a distinct band at an approximate molecular weight of 26 kDa in immunoblotting in reaction to an anti-G1 mouse serum, indicated that G1 protein was successfully expressed by this recombinant construct in the host cells.

Keywords

Bovine ephemeral fever virus (BEFV)

Recombinant G1 protein

DNA vaccine

Subjects